Light sensitive compounds for instant determination of organ function

ABSTRACT

Highly hydrophilic indole and benzoindole derivatives that absorb and fluoresce in the visible region of light are disclosed. These compounds are useful for physiological and organ function monitoring. Particularly, the molecules of the invention are useful for optical diagnosis of renal and cardiac diseases and for estimation of blood volume in vivo.

[0001] This application is a Continuation-In-Part of U.S. patentapplication Ser. No. 09/688,942 filed Oct. 16, 2000, now pending andexpressly incorporated by reference herein in its entirety.

FIELD OF THE INVENTION

[0002] This invention relates to novel optical probes for use inphysiological function monitoring, particularly indole and benzoindolecompounds.

BACKGROUND OF THE INVENTION

[0003] Dynamic monitoring of physiological functions of patients at thebedside is highly desirable in order to minimize the risk of acute renalfailure brought about by various clinical, physiological, andpathological conditions (C. A. Rabito, L. S. T. Fang, and A. C. Waltman,Renal function in patients at risk with contrast material-induced acuterenal failure: Noninvasive real-time monitoring, Radiology 1993, 186,851-854; N. L. Tilney, and J. M. Lazarus, Acute renal failure insurgical patients: Causes, clinical patterns, and care, Surgical Clinicsof North America, 1983, 63, 357-377; B. E. VanZee, W. E. Hoy, and J. R.Jaenike, Renal injury associated with intravenous pyelography innon-diabetic and diabetic patients, Annals of Internal Medicine, 1978,89, 51-54; S. Lundqvist, G. Edbom, S. Groth, U. Stendahl, and S. O.Hietala, Iohexol clearance for renal function measurement in gynecologiccancer patients, Acta Radiologica, 1996, 37, 582-586; P. Guesry, L.Kaufman, S. Orlof, J. A. Nelson, S. Swann, and M. Holliday Measurementof glomerular filtration rate by fluorescent excitation ofnon-radioactive meglumine iothalamate, Clinical Nephrology, 1975, 3,134-138). This monitoring is particularly important in the case ofcritically ill or injured patients because a large percentage of thesepatients face the risk of multiple organ failure (MOF), resulting indeath (C. C. Baker et al., Epidemiology of Trauma Deaths, AmericanJournal of Surgery, 1980, 144-150; G. Regel et al., Treatment Results ofPatients with Multiple Trauma: An Analysis of 3406 Cases Treated Between1972 and 1991 at a German Level I Trauma Center, Journal of Trauma,1995, 38, 70-77). MOF is a sequential failure of lung, liver, andkidneys, and is incited by one or more severe causes such as acute lunginjury (ALI), adult respiratory distress syndrome (ARDS),hypermetabolism, hypotension, persistent inflammatory focus, or sepsissyndrome. The common histological features of hypotension and shockleading to MOF include tissue necrosis, vascular congestion,interstitial and cellular edema, hemorrhage, and microthrombi. Thesechanges affect the lung, liver, kidneys, intestine, adrenal glands,brain, and pancreas, in descending order of frequency (J. Coalson,Pathology of Sepsis, Septic Shock, and Multiple Organ Failure, NewHorizons: Multiple Organ Failure Syndrome, D. J. Bihari and F. B. Cerra(Eds.), Society of Critical Care Medicine, Fullerton, Calif., 1986, pp.27-59). The transition from early stages of trauma to clinical MOF ismarked by the extent of liver and renal failure and a change inmortality risk from about 30% to about 50% (F. B. Cerra, Multiple OrganFailure Syndrome. New Horizons: Multiple Organ Failure, D. J. Bihari andF. B. Cerra (Eds). Society of Critical Care Medicine, Fullerton, Calif.,1989, pp. 1-24).

[0004] Serum creatinine measured at frequent intervals by clinicallaboratories is currently the most common way of assessing renalfunction and following the dynamic changes in renal function which occurin critically ill patients (P. D. Doolan, E. L. Alpen, and G. B. Theil,A clinical appraisal of the plasma concentration and endogenousclearance of creatinine, American Journal of Medicine, 1962, 32, 65-79;J. B. Henry (Ed). Clinical Diagnosis and Management by LaboratoryMethods. 17th Edition, W. B. Saunders, Philadelphia, Pa., 1984); C. E.Speicher, The right test: A physician's guide to laboratory medicine, W.B. Saunders, Philadelphia, Pa., 1989). These values are frequentlymisleading, since age, state of hydration, renal perfusion, muscle mass,dietary intake, and many other clinical and anthropometric variablesaffect the value. In addition, a single value returned several hoursafter sampling is difficult to correlate with other importantphysiologic events such as blood pressure, cardiac output, state ofhydration and other specific clinical events (e.g., hemorrhage,bacteremia, ventilator settings and others). An approximation ofglomerular filtration rate can be made via a 24-hour urine collection,but this requires 24 hours to collect the sample, several more hours toanalyze the sample, and a meticulous bedside collection technique. Newor repeat data are equally cumbersome to obtain. Occasionally, changesin serum creatinine must be further adjusted based on the values forurinary electrolytes, osmolality, and derived calculations such as the“renal failure index” or the “fractional excretion of sodium.” Theserequire additional samples of serum collected contemporaneously withurine samples and, after a delay, precise calculations. Frequently,dosing of medication is adjusted for renal function and thus can beequally as inaccurate, equally delayed, and as difficult to reassess asthe values upon which they are based. Finally, clinical decisions in thecritically ill population are often as important in their timing as theyare in their accuracy.

[0005] Exogenous markers such as inulin, iohexol, ⁵¹Cr-EDTA, Gd-DTPA, or^(99m)Tc-DTPA have been reported to measure the glomerular filtrationrate (GFR) (P. L. Choyke, H. A. Austin, and J. A. Frank, Hydratedclearance of gadolinium-DTPA as a measurement of glomerular filtrationrate, Kidney International, 1992, 41, 1595-1598; M. F. Tweedle, X.Zhang, M. Fernandez, P. Wedeking, A. D. Nunn, and H. W. Strauss, Anoninvasive method for monitoring renal status at bedside, Invest.Radiol., 1997, 32, 802-805; N. Lewis, R. Kerr, and C. Van Buren,Comparative evaluation of urographic contrast media, inulin, and ^(99m)Tc-DTPA clearance methods for determination of glomerular filtrationrate in clinical transplantation, Transplantation, 1989, 48, 790-796).Other markers such as ¹²³I and ¹²⁵I labeled o-iodohippurate or^(99m)Tc-MAG₃ are used to assess tubular secretion process (W. N. Tauxe,Tubular Function, in Nuclear Medicine in Clinical Urology andNephrology, W. N. Tauxe and E. V. Dubovsky, Editors, pp. 77-105,Appleton Century Crofts, East Norwalk, 1985; R. Muller-Suur, and C.Muller-Suur, Glomerular filtration and tubular secretion of MAG ₃ in ratkidney, Journal of Nuclear Medicine, 1989, 30, 1986-1991). However,these markers have several undesirable properties such as the use ofradioactivity or ex-vivo handling of blood and urine samples. Thus, inorder to assess the status and to follow the progress of renal disease,there is a considerable interest in developing a simple, safe, accurate,and continuous method for determining renal function, preferably bynon-radioactive procedures. Other organs and physiological functionsthat would benefit from real-time monitoring include the heart, theliver, and blood perfusion, especially in organ transplant patients.

[0006] Hydrophilic, anionic substances are generally recognized to beexcreted by the kidneys (F. Roch-Ramel, K. Besseghir, and H. Murer,Renal excretion and tubular transport of organic anions and cations,Handbook of Physiology, Section 8, Neurological Physiology, Vol. II, E.E. Windhager, Editor, pp. 2189-2262, Oxford University Press, New York,1992; D. L. Nosco, and J. A. Beaty-Nosco, Chemistry of technetiumradiopharmaceuticals 1: Chemistry behind the development oftechnetium-^(99m) compounds to determine kidney function, CoordinationChemistry Reviews, 1999, 184, 91-123). It is further recognized thatdrugs bearing sulfonate residues exhibit improved clearance through thekidneys (J. Baldas, J. Bonnyman, Preparation, HPLC studies andbiological behavior of techentium-^(99m) and 99^(m)TcN0-radiopharmaceuticals based on quinoline type ligands, Nuc. Med.Biol., 1999, 19, 491-496; L. Hansen, A. Taylor, L., L. G. Marzilli,Synthesis of the sulfonate and phosphonate derivatives ofmercaptoacetyltriglycine. X-ray crystal structure of Na ₂[ReO(mercaptoacetylglycylglycylaminomethane-sulfonate)]3H₂O, Met.-BasedDrugs, 1994, 1, 31-39).

[0007] Assessment of renal function by continuously monitoring the bloodclearance of exogenous optical markers, viz., fluorescein bioconjugatesderived from anionic polypeptides, has been developed by us and byothers (R. B. Dorshow, J. E. Bugaj, B. D. Burleigh, J. R. Duncan, M. A.Johnson, and W. B. Jones, Noninvasive fluorescence detection of hepaticand renal function, Journal of Biomedical Optics, 1998, 3, 340-345; M.Sohtell et al., FITC-Inulin as a Kidney Tubule Marker in the Rat, Acta.Physiol. Scand., 1983, 119, 313-316, each of which is expresslyincorporated herein by reference). The main drawback of high molecularweight polypeptides is that they are immunogenic. In addition, largepolymers with narrow molecular weight distribution are difficult toprepare, especially in large quantities. Thus, there is a need in theart to develop low molecular weight compounds that absorb and/or emitlight that can be used for assessing renal, hepatic, cardiac and otherorgan functions.

SUMMARY OF THE INVENTION

[0008] The present invention overcomes these difficulties byincorporating hydrophilic anionic or polyhydroxy residues in the form ofsulfates, sulfonates, sulfamates, phosphates, polyethers, polyaminocarboxylic acids and their derivatives, and strategically positionedhydroxyl groups. Thus, the present invention is related to novel dyescontaining multiple hydrophilic moieties and their use as diagnosticagents for assessing organ function and functional status of tumors.

[0009] The novel compounds of the present invention comprise dyes ofFormulas 1 to 6 which are hydrophilic and absorb light in the visibleand near infrared regions of the electromagnetic spectrum. The bloodclearance rate can be modified by formulating the dyes in liposomes,micelles, or other microparticles. This enhances their use forphysiological monitoring of many organs. The ease of modifying theclearance pathways of the dyes after in vivo administration permitstheir use for physiological monitoring. Compounds with longer bloodpersistence are useful for angiography and organ perfusion analysis,which is particularly useful in organ transplant and critical illpatients. Predominant kidney clearance of the dyes enables their use fordynamic renal function monitoring, and rapid liver uptake of the dyesfrom blood serves as a useful index for the evaluation of hepaticfunction.

[0010] As illustrated in FIGS. 1-7, these dyes are designed to inhibitaggregation in solution by preventing intramolecular and intermolecularinduced hydrophobic interactions.

[0011] The present invention relates particularly to the novel compoundscomprising indoles of the general Formula 1

[0012] wherein R₃, R4, R₅, R₆, and R₇, and Y₁ are independently selectedfrom the group consisting of —H, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl,C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl, glucose derivatives ofR groups, saccharides, amino, C1-C10 aminoalkyl, cyano, nitro, halogen,hydrophilic peptides, arylpolysulfonates, C1-C10 alkyl, C1-C10 aryl,—SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂9-(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCON H(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCON H(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b)) —CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₁ is selected fromthe group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se; a, b,d, f, h, i, and j independently vary from 1-10; c, e, g, and kindependently vary from 1-100; R_(a), R_(b), R_(c), and R_(d) aredefined in the same manner as Y₁; T is either H or a negative charge.

[0013] The present invention also relates to the novel compoundscomprising benzoindoles of general Formula 2

[0014] wherein R₈, R₉, R₁₀, R₁₁, R₁₂, R₁₃, R₁₄, and Y₂ are independentlyselected from the group consisting of —H, C1-C10 alkoxyl, C1-C10polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl,glucose derivatives of R groups, saccharides, amino, C1-C10 aminoalkyl,cyano, nitro, halogen, hydrophilic peptides, arylpolysulfonates, C1-C10alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, (CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CON H(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NH PO₃HT, —(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂,—(CH₂)_(a)OCO(CH₂)_(b)PO₃HT, —(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT, —(CH₂)_(a)CON H(CH₂)_(b)PO₃T₂,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂,—(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and —(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂,—CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₂ is selected fromthe group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se; a, b,d, f, h, i, and j independently vary from 1-10; c, e, g, and kindependently vary from 1-100; R_(a), R_(b), R_(c), and R_(d) aredefined in the same manner as Y₂; T is either H or a negative charge.

[0015] The present invention also relates to the novel compoundscomprising cyanine dyes of general Formula 3

[0016] wherein R₁₅, R₁₆, R₁₇, R₁₈, R₁₉, R₂₀, R₂₁, R₂₂, R₂₃, Y₃, and Z₃are independently selected from the group consisting of —H, C1-C10alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20polyhydroxyaryl, glucose derivatives of R groups, saccharides, amino,C1-C10 aminoalkyl, cyano, nitro, halogen, hydrophilic peptides,arylpolysulfonates, C1-C10 alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—CH₂)_(a)OCON H(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCON H(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₃ and X₃ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₃ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); a, b, d, f, h, i, and j independently vary from1-10; c, e, g, and k independently vary from 1-100; a₃ and b₃ vary from0 to 5; R_(a), R_(b), R_(c), and R_(d) are defined in the same manner asY₃; T is either H or a negative charge.

[0017] The present invention further relates to the novel compoundscomprising cyanine dyes of general Formula 4

[0018] wherein R₂₄, R₂₅, R₂₆, R₂₇, R₂₈, R₂₉, R₃₀, R₃₁, R₃₂, R₃₃, R₃₄,R₃₅, R₃₆, Y₄, and Z₄ are independently selected from the groupconsisting of —H, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20polyhydroxyalkyl, C5-C20 polyhydroxyaryl, glucose derivatives of Rgroups, saccharides, amino, C1-C10 aminoalkyl, cyano, nitro, halogen,hydrophilic peptides, arylpolysulfonates, C1-C10 alkyl, C1-C10 aryl,—SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—N H₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₄ and X4 are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₄ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); a₄ and b₄ vary from 0 to 5; a, b, d, f, h, i,and j independently vary from 1-10; c, e, g, and k independently varyfrom 1-100; R_(a), R_(b), R_(c), and R_(d) are defined in the samemanner as Y₄; T is either H or a negative charge.

[0019] The present invention also relates to the novel compoundscomprising cyanine dyes of general Formula 5

[0020] wherein R₃₇, R₃₈, R₃₉, R₄₀, R₄₁, R₄₂, R₄₃, R₄₄, R₄₅, Y₅, and Z₅are independently selected from the group consisting of —H, C1-C10alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20polyhydroxyaryl, glucose derivatives of R groups, saccharides, amino,C1-C10 aminoalkyl, cyano, nitro, halogen, hydrophilic peptides,arylpolysulfonates, C1-C10 alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCON H(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCON H(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₅ and X₅ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₅ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); D5 is a single or a double bond; A₅, B₅ and E₅may be the same or different and are selected from the group consistingof —O—, —S—, —Se—, —P—, —NR_(a), —CR_(c)R_(d), CR_(c), alkyl, and —C═O;A₅, B₅, D₅, and E₅ may together form a 6 or 7 membered carbocyclic ringor a 6 or 7 membered heterocyclic ring optionally containing one or moreoxygen, nitrogen, or a sulfur atom; a, b, d, f, h, i, and jindependently vary from 1-10; c, e, g, and k independently vary from1-100; a₅ and b₅ vary from 0 to 5; R_(a), R_(b), R_(c), and R_(d) aredefined in the same manner as Y₅; T is either H or a negative charge.

[0021] The present invention also relates to the novel compoundscomprising cyanine dyes of general Formula 6

[0022] wherein R₄₆, R₄₇, R₄₉, R₄₉, R₅₀, R₅₁, R₅₂, R₅₃, R₅₄, R₅₅, R₅₆,R₅₇ and R₅₈, Y₆, and Z₆ are independently selected from the groupconsisting of —H, C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20polyhydroxyalkyl, C₅-C₂₀ polyhydroxyaryl, glucose derivatives of Rgroups, saccharides, amino, C1-C10 aminoalkyl, cyano, nitro, halogen,hydrophilic peptides, arylpolysulfonates, C1-C10 alkyl, C1-C10 aryl,—SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(r)N H₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—N H₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₆ and X₆ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₆ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); D6 is a single or a double bond; A6, B₆ and E₆may be the same or different and are selected from the group consistingof —O—, —S—, —Se—, —P—, —NR_(a), —CR_(c)R_(d), CR_(c), alkyl, and —C═O;A₆, B₆, D₆, and E₆ may together form a 6 or 7 membered carbocyclic ringor a 6 or 7 membered heterocyclic ring optionally containing one or moreoxygen, nitrogen, or sulfur atom; a, b, d, f, h, i, and j independentlyvary from 1-10; c, e, g, and k independently vary from 1-100; a₆ and b₆vary from 0 to 5; R_(a), R_(b), R_(c), and R_(d) are defined in the samemanner as Y₆; T is either H or a negative charge.

[0023] A hydrophilic chelate such as ethylenediaminetetraacetic acid(EDTA), diethylenetriaminepentaacetic acid (DPTA),1,4,7,10-tetraazacyclododecanetetraacetic acid (DOTA), or theirderivatives, can be attached to the compounds of Formulas 1-6 as one ormore R groups. These structures are expected to be highly water soluble.

[0024] The inventive compounds, compositions, and methods areadvantageous since they provide a real-time, accurate, repeatablemeasure of renal excretion rate using exogenous markers under specificyet changing circumstances. This represents a substantial improvementover any currently available or widely practiced method, sincecurrently, no reliable, continuous, repeatable bedside method for theassessment of specific renal function by optical methods exists.Moreover, since the inventive method depends solely on the renalelimination of the exogenous chemical entity, the measurement isabsolute and requires no subjective interpretation based on age, musclemass, blood pressure, etc. In fact it represents the nature of renalfunction in a particular patient, under these particular circumstances,at a precise moment in time.

[0025] The inventive compounds, compositions, and methods providesimple, efficient, and effective monitoring of organ function. Thecompound is administered and a sensor, either external or internal, isused to detect absorption and/or emission to determine the rate at whichthe compound is cleared from the blood. By altering the R groups, thecompounds may be rendered more organ specific.

BRIEF DESCRIPTION OF THE DRAWINGS

[0026]FIG. 1: Reaction pathway for the preparation of indolederivatives.

[0027]FIG. 2: Reaction pathway for the preparation of benzoindolederivatives.

[0028]FIG. 3: Reaction pathway for the preparation of indocarbocyaninederivatives.

[0029]FIG. 4: Reaction pathway for the preparation ofbenzoindocarbocyanine derivatives.

[0030]FIG. 5: Reaction pathway for the preparation of robustindocarbocyanine derivatives.

[0031]FIG. 6: Reaction pathway for the preparation of robustbenzoindocarbocyanine derivatives.

[0032]FIG. 7: Reaction pathway for the preparation of long-wavelengthabsorbing indocarbocyanine derivatives.

[0033]FIG. 8A: Absorption spectrum of indoledisulfonate in water.

[0034]FIG. 8B: Emission spectrum of indoledisulfonate in water.

[0035]FIG. 9A: Absorption spectrum of indocarbocyaninetetrasulfonate inwater.

[0036]FIG. 9B: Emission spectrum of indocarbocyaninetetrasulfonate inwater.

[0037]FIG. 10A: Absorption spectrum of chloroindocarbocyanine inacetonitrile.

[0038]FIG. 10B: Emission spectrum of chloroindocarbocyanine inacetonitrile.

[0039]FIG. 11: Blood clearance profile of carbocyanine-polyaspartic (10kDa) acid conjugate in a rat.

[0040]FIG. 12: Blood clearance profile of carbocyanine-polyaspartic (30kDa) acid conjugate in a rat.

[0041]FIG. 13: Blood clearance profile of indoledisulfonate in a rat.

[0042]FIG. 14: Blood clearance profile of carbocyaninetetrasulfonates ina rat.

DETAILED DESCRIPTION

[0043] In one embodiment the dyes of the invention serve as probes forcontinuous monitoring of renal function, especially for critically illpatients and kidney transplant patients.

[0044] In another embodiment, the dyes of the invention are useful fordynamic hepatic function monitoring, especially for critically illpatients and liver transplant patients.

[0045] In another embodiment, the dyes of the invention are useful forreal-time determination of cardiac function, especially in patients withcardiac diseases.

[0046] In another embodiment, the dyes of the invention are useful formonitoring organ perfusion, especially for critically ill, cancer, andorgan transplant patients.

[0047] In another embodiment, the dyes are useful for assessing thefunctional status of tumors and for monitoring tumor perfusion, such asin renal or hepatic cancer patients.

[0048] The novel dyes of the present invention are prepared according tothe methods well known in the art, as illustrated in general in FIGS.1-7 and described for specific compounds in Examples 1-11.

[0049] In one embodiment, the novel compounds, also called tracers, ofthe present invention have the Formula 1, wherein R₃, R₄, R₅, R₆ and R₇,and Y₁ are independently selected from the group consisting of —H, C1-C5alkoxyl, C1-C5 polyalkoxyalkyl, C1-C10 polyhydroxyalkyl, C5-C20polyhydroxyaryl, mono- and disaccharides, nitro, hydrophilic peptides,arylpolysulfonates, C1-C5 alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—N H₂, —(CH₂)_(h) —N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₁ is selected fromthe group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se; a, b,d, f, h, l, and j independently vary from 1-5; c, e, g, and kindependently vary from 1-20; R_(a), R_(b), R_(c), and R_(d) are definedin the same manner as Y₁; T is a negative charge.

[0050] In another embodiment, the novel compounds of the presentinvention have the general Formula 2, wherein R₈, R₉, R₁₀, R₁₁, R₁₂,R₁₃, R₁₄, and Y₂ are independently selected from the group consisting of—H, C1-C5 alkoxyl, C1-C5 polyalkoxyalkyl, C1-C10 polyhydroxyalkyl,C5-C20 polyhydroxyaryl, mono- and disaccharides, nitro, hydrophilicpeptides, arylpolysulfonates, C1-C5 alkyl, C1-C10 aryl, —SO₃T, —CO₂T,—OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—CH₂(CH₂—C—CH₂)C—CH₂—OH, —(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T,—(CH₂)_(r)N H₂, —CH₂—(CH₂—O—CH₂)_(g)—CH₂—N H₂,—(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₂ is selected fromthe group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se; a, b,d, f, h, l, and j independently vary from 1-5; c, e, g, and kindependently vary from 1-20; R_(a), R_(b), R_(c), and R_(d) are definedin the same manner as Y₂; T is a negative charge.

[0051] In another embodiment, the novel compositions of the presentinvention have the general Formula 3, wherein R₁₅, R₁₆, R₁₇, R₁₈, R₁₉,R₂₀, R₂₁, R₂₂, R₂₃, Y₃, and Z₃ are independently selected from the groupconsisting of —H, C1-C5 alkoxyl, C1-C5 polyalkoxyalkyl, C1-C10polyhydroxyalkyl, C5-C20 polyhydroxyaryl, mono- and disaccharides,nitro, hydrophilic peptides, arylpolysulfonates, C1-C5 alkyl, C1-C10aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₃ and X₃ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₃ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); a, b, d, f, h, i, and j independently vary from1-5; c, e, g, and k independently vary from 1-50; a₃ and b₃ vary from 0to 5; R_(a), R_(b), R_(c), and R_(d) are defined in the same manner asY₃; T is either H or a negative charge.

[0052] In another embodiment, the novel compounds of the presentinvention have the general Formula 4, wherein R₂₄, R₂₅, R₂₆, R₂₇, R₂₈,R₂₉, R₃₀, R₃₁, R₃₂, R₃₃, R₃₄, R₃₅, R₃₆, Y₄, and Z₄ are independentlyselected from the group consisting of —H, C1-C5 alkoxyl, C1-C5polyalkoxyalkyl, C1-C10 polyhydroxyalkyl, C5-C20 polyhydroxyaryl, mono-and disaccharides, nitro, hydrophilic peptides, arylpolysulfonates,C1-C5 alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T,—(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—N H₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₄ and X4 are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₄ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); a₄ and b₄ vary from 0 to 5; a, b, d, f, h, i,and j independently vary from 1-5; c, e, g, and k independently varyfrom 1-50; R_(a), R_(b), R_(c), and R_(d) are defined in the same manneras Y₄; T is either H or a negative charge.

[0053] In another embodiment, the novel compounds of the presentinvention have the general Formula 5, wherein R₃₇, R₃₈, R₃₉, R₄₀, R₄₁,R₄₂, R₄₃, R₄₄, R₄₅, Y₅, and Z₅ are independently selected from the groupconsisting of —H, C1-C5 alkoxyl, C1-C5 polyalkoxyalkyl, C1-C10polyhydroxyalkyl, C5-C20 polyhydroxyaryl, mono- and disaccharides,nitro, hydrophilic peptides, arylpolysulfonates, C1-C5 alkyl, C1-C10aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₅ and X₅ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₅ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a) D₅ is a single or a double bond; A₅, B₅ and E₅may be the same or different and are selected from the group consistingof —O—, —S—, —NR_(a), —CR_(c)R_(d), CR_(c), and alkyl; A₅, B₅, D₅, andE₅ may together form a 6 or 7 membered carbocyclic ring or a 6 or 7membered heterocyclic ring optionally containing one or more oxygen,nitrogen, or sulfur atom; a, b, d, f, h, i, and j independently varyfrom 1-5; c, e, g, and k independently vary from 1-50; a₅ and b₅ varyfrom 0 to 5; R_(a), R_(b), R_(c), and R_(d) are defined in the samemanner as Y₅; T is either H or a negative charge.

[0054] In yet another embodiment, the novel compounds of the presentinvention have the general Formula 6, wherein R₄₆, R₄₇, R₄₈, R₄₉, R₅₀,R₅₁, R₅₂, R₅₃, R₅₄, R₅₅, R₅₆, R₅₇, R₅₈, Y₆, and Z₆ are independentlyselected from the group consisting of —H, C1-C5 alkoxyl, C1-C5polyalkoxyalkyl, C1-C10 polyhydroxyalkyl, C5-C20 polyhydroxyaryl, mono-and disaccharides, nitro, hydrophilic peptides, arylpolysulfonates,C1-C5 alkyl, C1-C10 aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T,—(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH, —(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; W₆ and X₆ are selectedfrom the group consisting of —CR_(c)R_(d), —O—, —NR_(c), —S—, and —Se;V₆ is a single bond or is selected from the group consisting of —O—,—S—, —Se—, and —NR_(a); D₆ is a single or a double bond; A₆, B₆ and E₆may be the same or different and are selected from the group consistingof —O—, —S—, —NR_(a), —CR_(c)R_(d), CR_(c), and alkyl; A, B₆, D₆, and E₆may together form a 6 or 7 membered carbocyclic ring or a 6 or 7membered heterocyclic ring optionally containing one or more oxygen,nitrogen, or sulfur atom; a, b, d, f, h, i, and j independently varyfrom 1-5; c, e, g, and k independently vary from 1-50; a₅ and b₅ varyfrom 0 to 5; R_(a), R_(b), R_(c), and R_(d) are defined in the samemanner as Y₆; T is either H or a negative charge.

[0055] The compounds of the invention can be formulated into diagnosticand therapeutic compositions for enteral or parenteral administration.These compositions contain an effective amount of the dye along withconventional pharmaceutical carriers and excipients appropriate for thetype of administration contemplated. For example, parenteralformulations advantageously contain the inventive agent in a sterileaqueous solution or suspension. Parenteral compositions may be injecteddirectly or mixed with a large volume parenteral composition forsystemic administration. Such solutions also may containpharmaceutically acceptable buffers and, optionally, electrolytes suchas sodium chloride.

[0056] Formulations for enteral administration may vary widely, as iswell known in the art. In general, such formulations are liquids, whichinclude an effective amount of the inventive agent in aqueous solutionor suspension. Such enteral compositions may optionally include buffers,surfactants, thixotropic agents, and the like. Compositions for oraladministration may also contain flavoring agents and other ingredientsfor enhancing their organoleptic qualities.

[0057] The compositions are administered in doses effective to achievethe desired effect or result. The dosage of the tracers may varyaccording to the clinical procedure contemplated and generally rangesfrom 1 picomolar to 100 millimolar. The compositions may be administeredto a patient, typically a warm-blooded animal either systemically orlocally to the organ or tissue to be imaged, and the patient thensubject to the imaging procedure. The tracers may be administered to thepatient by any suitable method, including intravenous, intraperitoneal,or subcutaneous injection or infusion, oral administration, transdermalabsorption through the skin, aerosols, or by inhalation. The detectionof the tracers is achieved by optical fluorescence, absorbance, or lightscattering methods known in the art (Muller et al. Eds., Medical OpticalTomography, SPIE Volume IS11, 1993, which is expressly incorporatedherein by reference) using invasive or non-invasive probes such asendoscopes, catheters, ear clips, hand bands, surface coils, fingerprobes, and the like. Physiological function is correlated with theclearance profiles and rates of these agents from body fluids (R. B.Dorshow et al., Non-Invasive Fluorescence Detection of Hepatic and RenalFunction, Bull. Am. Phys. Soc. 1997, 42, 681, which is expresslyincorporated by reference herein).

[0058] The inventive composition may be administered for imaging by morethan one modality. As one example, the composition may be used forimaging by optical imaging alone, by nuclear imaging alone, or by bothoptical and nuclear imaging modalities when a radioactive isotope isincluded in the chemical formula, such as replacing a halogen atom witha radioactive halogen, and/or including a radioactive metal ion such asTc⁹⁹, In¹¹¹, etc. As another example, the composition may be used forimaging by optical imaging alone, by magnetic resonance (MR) alone, orby both optical and MR modalities when a paramagnetic metal ion such asgadolinium or manganese is included in the chemical formula.

[0059] It will also be appreciated that the inventive compositions maybe administered with other contrast agents or media used to enhance animage from a non-optical modality. These include agents for enhancing animage obtained by modalities including but not limited to MR, ultrasound(US), x-ray, positron emission tomography (PET), computed tomography(CT), single photon emission computed tomography (SPECT), optoacoustic(e.g. U.S. Pat. Nos. 5,840,023 and 5,977,538 which are expresslyincorporated by reference herein in their entirety), etc. Both opticaland non-optical agents may be formulated as a single composition (thatis, one composition containing one, two, or more components, forexample, an optical agent and a MR agent), or may be formulated asseparate compositions. The inventive optical imaging contrast agent andthe non-optical contrast agent are administered in doses effective toachieve the desired enhancement, diagnosis, therapy, etc., as known toone skilled in the art. The inventive compositions, either alone orcombined with a contrast agent, may be administered to a patient,typically a warm-blooded animal, systemically or locally to the organ ortissue to be imaged. The patient is then imaged by optical imagingand/or by another modality. As one example of this embodiment, theinventive compounds may be added to contrast media compositions. Asanother example, the inventive compositions may be co-administered withcontrast media, either simultaneously or within the same diagnosticand/or therapeutic procedure (for example, administering the inventivecomposition and administering a contrast agent then performing opticalimaging followed by another imaging modality, or administering theinventive composition and administering a contrast agent then performinganother imaging modality followed by optical imaging, or administeringthe inventive composition and optical imaging, then administering acontrast agent and MR, US, CT, etc. imaging, or administering a contrastagent and imaging by MR, US, CT, etc., then administering the inventivecomposition and optical imaging, or administering the inventivecomposition and a contrast agent, and simultaneously imaging by anoptical modality and MR, US, CT, etc.). As another example, an opticalimaging agent may be added as an additive or excipient for a non-opticalimaging modality. In this embodiment, the optically active component,such as the dyes disclosed herein, could be added as a buffering agentto control pH or as a chelate to improve formulation stability, etc. inMR contrast media, CT contrast media, x-ray contrast media, US contrastmedia, etc. The MR, CT, x-ray, US contrast media would then alsofunction as an optical imaging agent. The information obtained from themodality using the non-optical contrast agent is useful in combinationwith the image obtained using the optical contrast agent.

[0060] In one embodiment, the agents may be formulated as micelles,liposomes, microcapsules, or other microparticles. These formulationsmay enhance delivery, and localization of the inventive compounds to/atthe desired organ or site. The target specificity of these formulationscan be enhanced by using suitable targeting molecules such as peptides,saccharides, fatty acids, etc. Preparation and loading of these are wellknown in the art.

[0061] As one example, liposomes may be prepared from dipalmitoylphosphatidylcholine (DPPC) or egg phosphatidylcholine (PC) because thislipid has a low heat transition. Liposomes are made using standardprocedures as known to one skilled in the art (e.g., Braun-Falco et al.,(Eds.), Griesbach Conference, Liposome Dermatics, Springer-Verlag,Berlin (1992)). Polycaprolactone, poly(glycolic) acid, poly(lactic)acid, polyanhydride or lipids may be formulated as microspheres. As anillustrative example, the optical agent may be mixed with polyvinylalcohol (PVA), the mixture then dried and coated with ethylene vinylacetate, then cooled again with PVA. In a liposome, the optical agentmay be within one or both lipid bilayers, in the aqueous between thebilayers, or with the center or core. Liposomes may be modified withother molecules and lipids to form a cationic liposome. Liposomes mayalso be modified with lipids to render their surface more hydrophilicwhich increases their circulation time in the bloodstream. Thethus-modified liposome has been termed a “stealth” liposome, or along-lived liposome, as described in U.S. Pat. Nos. 6,277403; 6,610,322;5,631,018; 5,395,619; and 6,258,378, each of which is expresslyincorporated by reference herein in its entirety, and in StealthLiposomes, Lasic and Martin (Eds.) 1995, CRC Press, London.Encapsulation methods include detergent dialysis, freeze drying, filmforming, injection, as known to one skilled in the art and disclosed in,for example, U.S. Pat. No. 6,406,713 which is expressly incorporated byreference herein in its entirety.

[0062] The agent formulated in liposomes, microcapsules, etc. may beadministered by any of the routes previously described. In a formulationapplied topically, the optical agent is slowly released over time. In aninjectable formulation, the liposome capsule circulates in thebloodstream and is delivered to a desired site.

[0063] As another example, microparticles such as ultra small iron oxideparticles (USPIO) and other metallic particles such as silver or goldparticles coated with or attached (covalently or non-covalently) withthe inventive compounds may be used for optical imaging and/or MRI. Suchparticles are known to one skilled in the art as disclosed in, forexample, U.S. Pat. No. 5,492,814 and Journal of Biomedical Optics 8(3),472-478 (July 2003) which are expressly incorporated by reference hereinin their entirety.

[0064] Organ function can be assessed either by the differences in themanner in which the normal and impaired cells remove the tracer from thebloodstream, by measuring the rate or accumulation of these tracers inthe organs or tissues, or by obtaining tomographic images of the organsor tissues. Blood pool clearance may be measured non-invasively fromconvenient surface capillaries such as those found in an ear lobe or afinger, for example, using an ear clip or finger clip sensor, or may bemeasured invasively using an endovascular catheter. Accumulation of thetracer within the cells of interest can be assessed in a similarfashion. The clearance of the tracer dyes may be determined by selectingexcitation wavelengths and filters for the emitted photons. Theconcentration-time curves may be analyzed in real time by amicroprocessor. In order to demonstrate feasibility of the inventivecompounds to monitor organ function, a non-invasive absorbance orfluorescence detection system to monitor the signal emanating from thevasculature infused with the compounds is used. Indole derivatives, suchas those of Formulas 1-6, fluoresce at a wavelength between 350 nm and1300 nm when excited at the appropriate wavelength as is known to, orreadily determined by, one skilled in the art.

[0065] In addition to the noninvasive techniques, a modified pulmonaryartery catheter can be used to make the necessary measurements (R. B.Dorshow, J. E. Bugaj, S. A. Achilefu, R. Rajagopalan, and A. H. Combs,Monitoring Physiological Function by Detection of Exogenous FluorescentContrast Agents, in Optical Diagnostics of Biological Fluids IV, A.Priezzhev and T. Asakura, Editors, Proceedings of SPIE 1999, 3599, 2-8,which is expressly incorporated by reference herein). Currently,pulmonary artery catheters measure only intravascular pressures, cardiacoutput and other derived measures of blood flow. Critically ill patientsare managed using these parameters, but rely on intermittent bloodsampling and testing for assessment of renal function. These laboratoryparameters represent discontinuous data and are frequently misleading inmany patient populations. Yet, importantly, they are relied upon heavilyfor patient assessment, treatment decisions, and drug dosing.

[0066] The modified pulmonary artery catheter incorporates an opticalsensor into the tip of a standard pulmonary artery catheter. Thiswavelength specific optical sensor can monitor the renal functionspecific elimination of an optically detectable chemical entity. Thus,by a method analogous to a dye dilution curve, real-time renal functioncan be monitored by the disappearance of the optically detectedcompound. Modification of a standard pulmonary artery catheter onlyrequires making the fiber optic sensor wavelength specific, as is knownto one skilled in this art. Catheters that incorporate fiber optictechnology for measuring mixed venous oxygen saturation currently exist.

[0067] The present invention may be used for rapid bedside evaluation ofrenal function and also to monitor the efficiency of hemodialysis. Theinvention is further demonstrated by the following examples. Since manymodifications, variations, and changes in detail may be made to thedescribed embodiments, it is intended that all matter in the foregoingdescription and shown in the accompanying drawings be interpreted asillustrative and not in a limiting sense.

Example 1 Synthesis of Indole Disulfonate (FIG. 1, Compound 5, Y₇=SO₃ ⁻;X₇=H: n=1)

[0068] A mixture of 3-methyl-2-butanone (25.2 mL), andp-hydrazinobenzenesulfonic acid (15 g) in acetic acid (45 mL) was heatedat 110° C. for 3 hours. After reaction, the mixture was allowed to coolto room temperature and ethyl acetate (100 mL) was added to precipitatethe product, which was filtered and washed with ethyl acetate (100 mL).The intermediate compound, 2,3,3-trimethylindolenium-5-sulfonate (FIG.1, compound 3) was obtained as a pink powder in 80% yield. A portion ofcompound 3 (9.2 g) in methanol (115 mL) was carefully added to asolution of KOH in isopropanol (100 mL). A yellow potassium salt of thesulfonate was obtained in 85% yield after vacuum-drying for 12 hours. Aportion of the 2,3,3-trimethylindolenium-5-sulfonate potassium salt (4g) and 1,3-propanesultone (2.1 g) was heated in dichlorobenzene (40 mL)at 110° C. for 12 hours. The mixture was allowed to cool to roomtemperature and the resulting precipitate was filtered and washed withisopropanol. The resulting pink powder was dried under vacuum to give97% of the desired compound.

[0069] Other compounds prepared by a similar method described aboveinclude polyhydroxyl indoles such as

Example 2 Synthesis of Indole Disulfonate (FIG. 1, Compound 5, Y₇=SO₃ ⁻:X₇=H; n=2)

[0070] This compound was prepared by the same procedure described inExample 1, except that 1,4-butanesultone was used in place of1,3-propanesultone.

Example 3 Synthesis of Benzoindole Disulfonate (FIG. 2, Compound 8, Y₇,Y₈=SO₃ ⁻; X₇=H; n=2)

[0071] This compound was prepared by the same procedure described inExample 1, except that hydrazinonaphthalenedisulfonic acid was used inplace of hydrazinobenzenesulfonic acid.

[0072] Other compounds prepared by a similar method includepolyhydroxyindoles such as:

Example 4 Synthesis of Benzoindole Disulfonate (FIG. 2, Compound 8, Y₇,Y₈=SO₃ ⁻; X₇=OH: n=4)

[0073] This compound was prepared by the same procedure described inExample 1, except that 3-hydroxymethyl-4-hydroxyl-2-butanone was used inplace of 3-methyl-2-butanone.

Example 5 Synthesis of Bis(ethylcarboxymethyl)indocyanine Dye

[0074]

[0075] A mixture of 1,1,2-trimethyl-[1H]-benz[e]indole (9.1 g, 43.58mmoles) and 3-bromopropanoic acid (10.0 g, 65.37 mmoles) in1,2-dichlorobenzene (40 mL) was heated at 110° C. for 12 hours. Thesolution was cooled to room temperature and the red residue obtained wasfiltered and washed with acetonitrile: diethyl ether (1:1) mixture. Thesolid obtained was dried under vacuum to give 10 g (64%) of light brownpowder. A portion of this solid (6.0 g; 16.56 mmoles), glutaconaldehydedianil monohydrochloride (2.36 g, 8.28 mmoles) and sodium acetatetrihydrate (2.93 g, 21.53 mmoles) in ethanol (150 mL) were refluxed for90 minutes. After evaporating the solvent, 40 mL of 2 N aqueous HCl wasadded to the residue and the mixture was centrifuged and the supernatantwas decanted. This procedure was repeated until the supernatant becamenearly colorless. About 5 mL of water:acetonitrile (3:2) mixture wasadded to the solid residue and lyophilized to obtain 2 g of dark greenflakes. The purity of the compound was established with ¹H-NMR andliquid chromatography/mass spectrometry (LC/MS).

Example 6 Synthesis of Bis(pentylcarboxymethyl)indocyanine Dye

[0076]

[0077] A mixture of 2,2,3-trimethyl-[1H]-benz[e]indole (20 g, 95.6mmoles) and 6-bromohexanoic acid (28.1 g, 144.1 mmoles) in1,2-dichlorobenzene (250 mL) was heated at 110° C. for 12 hours. Thegreen solution was cooled to room temperature and the brown solidprecipitate formed was collected by filtration. After washing the solidwith 1,2-dichlorobenzene and diethyl ether, the brown powder obtained(24 g, 64%) was dried under vacuum at room temperature. A portion ofthis solid (4.0 g; 9.8 mmoles), glutaconaldehyde dianilmonohydrochloride (1.4 g, 5 mmoles) and sodium acetate trihydrate (1.8g, 12.9 mmoles) in ethanol (80 mL) were refluxed for 1 hour. Afterevaporating the solvent, 20 mL of a 2 N aqueous HCl was added to theresidue and the mixture was centrifuged and the supernatant wasdecanted. This procedure was repeated until the supernatant becamenearly colorless. About 5 mL of water:acetonitrile (3:2) mixture wasadded to the solid residue and lyophilized to obtain about 2 g of darkgreen flakes. The purity of the compound was established with ¹H-NMR,HPLC, and LC-MS.

Example 7 Synthesis of polyhydroxyindole sulfonate (FIG. 3. Compound 13,Y₇, Y₈=O₃ ⁻; X₇═OH: n=2)

[0078] Phosphorus oxychloride (37 ml, 0.4 mole) was added dropwise withstirring to a cooled (−2° C.) mixture of dimethylformamide (DMF, 0.5mole, 40 mL) and dichloromethane (DCM, 40 mL), followed by the additionof acetone (5.8 g, 0.1 mole). The ice bath was removed and the solutionrefluxed for 3 hours. After cooling to room temperature, the product waseither partitioned in water/DCM, separated and dried, or was purified byfractional distillation. Nuclear magnetic resonance and mass spectralanalyses showed that the desired intermediate, 10, was obtained.Reaction of the intermediate with 2 equivalents of2,2,3-trimethyl-[H]-benz[e]indolesulfonate-N-propanoic acid and 2equivalents of sodium acetate trihydrate in ethanol gave a blue-greensolution after 1.5 hours at reflux. Further functionalization of the dyewith bis(isopropylidene)acetal protected monosaccharide is effected bythe method described in the literature (J. H. Flanagan, C. V. Owens, S.E. Romero, et al., Near infrared heavy-atom-modified fluorescent dyesfor base-calling in DNA-sequencing application using temporaldiscrimination. Anal. Chem., 1998, 70(13), 2676-2684).

Example 8 Synthesis of polyhydroxyindole sulfonate (FIG. 4. Compound 16,Y₇, Y₈=SO₃ ⁻: X₇—H; n=1)

[0079] Preparation of this compound was readily accomplished by the sameprocedure described in Example 6 using p-hydroxybenzenesulfonic acid inthe place of the monosaccharide, and benzoindole instead of indolederivatives.

Example 9 Synthesis of polyhydroxyindole Sulfonate (FIG. 5. Compound 20,Y₇, Y₈=H; X₇=OH: n=1)

[0080] The hydroxyindole compound was readily prepared by a literaturemethod (P. L. Southwick, J. G. Cairns, L. A. Ernst, and A. S. Waggoner,One pot Fischer synthesis of (2,3,3-trimethyl-3-H-indol-5-yl)-aceticacid derivatives as intermediates for fluorescent biolabels, Org. Prep.Proced. Int. Briefs, 1988, 20(3), 279-284). Reaction ofp-carboxymethylphenylhydrazine hydrochloride (30 mmol, 1 equiv.) and1,1-bis(hydroxymethyl)propanone (45 mmol, 1.5 equiv.) in acetic acid (50mL) at room temperature for 30 minutes and at reflux for 1 gave(3,3-dihydroxymethyl2-methyl-3-H-indol-5-yl)-acetic acid as a solidresidue.

[0081] The intermediate 2-chloro-1-formyl-3-hydroxymethylenecyclo-hexanewas prepared as described in the literature (G. A. Reynolds and K. H.Drexhage, Stable heptamethine pyrylium dyes that absorb in the infrared.J. Org. Chem., 1977, 42(5), 885-888). Equal volumes (40 mL each) ofdimethylformamide (DMF) and dichloromethane were mixed and the solutionwas cooled to −10° C. in acetone-dry ice bath. Under argon atmosphere,phosphorus oxychloride (40 mL) in dichloromethane was added dropwise tothe cool DMF solution, followed by the addition of 10 g ofcyclohexanone. The resulting solution was allowed to warm up to roomtemperature and heated at reflux for 6 hours. After cooling to roomtemperature, the mixture was poured into ice-cold water and stored at 4°C. for 12 hours. A yellow powder was obtained. Condensation of a portionof this cyclic dialdehyde (1 equivalent) with the indole intermediate (2equivalents) was carried out as described in Example 5. Further, thefunctionalization of the dye with bis (isopropylidene)acetal protectedmonosaccharide was effected by the method described in the literature(J. H. Flanagan, C. V. Owens, S. E. Romero, et al., Near infraredheavy-atom-modified fluorescent dyes for base-calling in DNA-sequencingapplication using temporal discrimination. Anal. Chem., 1998, 70(13),2676-2684).

Example 10 Synthesis of polyhydroxylbenzoindole Sulfonate (FIG. 6.Compound 22, Y₇, Y₈=H; X₇=OH; n=1)

[0082] A similar method described in Example 8 was used to prepare thiscompound by replacing the indole with benzoindole derivatives.

Example 11 Synthesis of Rigid Heteroatomic Indole Sulfonate (FIG. 7.Compound 27, Y₇, Y₈, X₇=H; n=1)

[0083] Starting with 3-oxo-4-cyclohexenone, this heteroatomichydrophilic dye was readily prepared as described in Example 8.

Example 12 Minimally Invasive Monitoring of the Blood Clearance Profileof the Dyes

[0084] A laser of appropriate wavelength for excitation of the dyechromophore was directed into one end of a fiber optic bundle and theother end was positioned a few millimeters from the ear of a rat. Asecond fiber optic bundle was also positioned near the same ear todetect the emitted fluorescent light, and the other end was directedinto the optics and electronics for data collection. An interferencefilter (IF) in the collection optics train was used to select emittedfluorescent light of the appropriate wavelength for the dye chromophore.

[0085] Sprague-Dawley or Fischer 344 rats were anesthetized withurethane administered via intraperitoneal injection at a dose of 1.35g/kg body weight. After the animals had achieved the desired plane ofanesthesia, a 21 gauge butterfly with 12″ tubing was placed in thelateral tail vein of each animal and flushed with heparinized saline.The animals were placed onto a heating pad and kept warm throughout theentire study. The lobe of the left ear was affixed to a glass microscopeslide to reduce movement and vibration.

[0086] Incident laser light delivered from the fiber optic was centeredon the affixed ear. Data acquisition was then initiated, and abackground reading of fluorescence was obtained prior to administrationof the test agent.

[0087] The compound was administered to the animal through a bolusinjection in the lateral tail vein. The dose was typically 0.05 to 20μmole/kg of body weight. The fluorescence signal rapidly increased to apeak value, then decayed as a function of time as the conjugate clearedfrom the bloodstream.

[0088] This procedure was repeated with several dye-peptide conjugatesin normal and tumored rats. Representative profiles are shown in FIGS.6-10.

[0089] While the invention has been disclosed by reference to thedetails of preferred embodiments of the invention, it is to beunderstood that the disclosure is intended in an illustrative ratherthan in a limiting sense, as it is contemplated that modifications willreadily occur to those skilled in the art, within the spirit of theinvention and the scope of the appended claims.

What is claimed is:
 1. A pharmaceutical composition comprising aneffective amount of the compound of formula 4

for a diagnostic or therapeutic procedure and a pharmaceuticallyacceptable carrier for administration to a mammal wherein at least oneof W₄ and X₄ is —CR_(c)R_(d) and the other is selected from the groupconsisting of —CR_(c)R_(d), —NR_(c), —O—, and —S—; R₂₄, R₂₅, R₂₆, R₂₇,R₂₈, R₂₉, R₃₀, R₃₁, R₃₂, R₃₃, R₃₄, R₃₅ and R₃₆, Y₄, and Z₄ areindependently selected from the group consisting of C1-C10 alkoxyl,C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl,glucose derivatives of R groups, saccharides, amino, C1-C10 aminoalkyl,cyano, nitro, halogen, hydrophilic peptides, arylpolysulfonates, C1-C10alkyl, C5-C20 aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —(CH₂)_(a)CON H(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCO(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T, —(CH₂)_(a)OCONH(CH₂)_(b)SO₃T,—(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂, —(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂,—(CH₂)_(a)NHPO₃HT, —(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b)) —CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; V₄ is a single bondor is selected from the group consisting of —O—, —S—, —Se—, and —NR_(a);a₄ and b₄ vary from 0 to 5; a, b, d, f, h, i, and j independently varyfrom 1-10; c, e, g, and k independently vary from 1-100; R_(a), R_(b),R_(c), and R_(d) are defined in the same manner as Y₄; and T is either Hor a negative charge.
 2. A pharmaceutical composition comprising aneffective amount of the compound of formula 4

for a diagnostic or therapeutic procedure and a pharmaceuticallyacceptable carrier for administration to a mammal wherein at least oneof W₄ and X₄ is 0 and the other is selected from the group consisting of—CR_(c)R_(d), —NR_(c), —O—, and —S—; R₂₄, R₂₅, R₂₆, R₂₇, R₂₈, R₂₉, R₃₀,R₃₁, R₃₂, R₃₃, R₃₄, R₃₅ and R₃₆, Y₄, and Z₄ are independently selectedfrom the group consisting of C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl,C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl, glucose derivatives ofR groups, saccharides, amino, C1-C10 aminoalkyl, cyano, nitro, halogen,hydrophilic peptides, arylpolysulfonates, C1-C10 alkyl, C5-C20 aryl,—SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCON H(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, -(CH₂)_(a)CON H(CH₂)_(b)PO₃HT,—(CH₂)_(a)CON H(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH—(CH₂)_(d)—CO₂T,—CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; V₄ is a single bond oris selected from the group consisting of —O—, —S—, —Se—, and —NR_(a); a₄and b₄ vary from 0 to 5; a, b, d, f, h, i, and j independently vary from1-10; c, e, g, and k independently vary from 1-100; R_(a), R_(b), R_(c),and R_(d) are defined in the same manner as Y₄; and T is either H or anegative charge.
 3. A pharmaceutical composition comprising an effectiveamount of the compound of formula 4

for a diagnostic or therapeutic procedure and a pharmaceuticallyacceptable carrier for administration to a mammal wherein at least oneof W₄ and X₄ is NR_(c) and the other is selected from the groupconsisting of —CR_(c)R_(d), —NR_(c), —O—, and —S—; R₂₄, R₂₅, R₂₆, R₂₇,R₂₈, R₂₉, R₃₀, R₃₁, R₃₂, R₃₃, R₃₄, R₃₅ and R₃₆, Y₄, and Z₄ areindependently selected from the group consisting of C1-C10 alkoxyl,C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl,glucose derivatives of R groups, saccharides, amino, C1-C10 aminoalkyl,cyano, nitro, halogen, hydrophilic peptides, arylpolysulfonates, C1-C10alkyl, C5-C20 aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —(CH₂)_(a)CONH(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCO(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T, —(CH₂)_(a)OCON H(CH₂)_(b)SO₃T,—(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂, —(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂,—(CH₂)_(a)NHPO₃HT, —(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT, —(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, (CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCON H(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCON H(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b)) —CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; V₄ is a single bondor is selected from the group consisting of —O—, —S—, —Se—, and —NR_(a);a₄ and b₄ vary from 0 to 5; a, b, d, f, h, i, and j independently varyfrom 1-10; c, e, g, and k independently vary from 1-100; R_(a), R_(b),R_(c), and R_(d) are defined in the same manner as Y₄; and T is either Hor a negative charge.
 4. A pharmaceutical composition comprising aneffective amount of the compound of formula 4

for a diagnostic or therapeutic procedure and a pharmaceuticallyacceptable carrier for administration to a mammal wherein at least oneof W₄ and X₄ is S and the other is selected from the group consisting of—CR_(c)R_(d), —NR_(c), —O—, and —S—; R₂₄, R₂₅, R₂₆, R₂₇, R₂₈, R₂₉, R₃₀,R₃₁, R₃₂, R₃₃, R₃₄, R₃₅ and R₃₆, Y₄, and Z₄ are independently selectedfrom the group consisting of C1-C10 alkoxyl, C1-C10 polyalkoxyalkyl,C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl, glucose derivatives ofR groups, saccharides, amino, C1-C10 aminoalkyl, cyano, nitro, halogen,hydrophilic peptides, arylpolysulfonates, C1-C10 alkyl, C5-C20 aryl,—SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T,—(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CON H(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCON H(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)₉—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b)) —CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; V₄ is a single bondor is selected from the group consisting of —O—, —S—, —Se—, and —NR_(a);a₄ and b₄ vary from 0 to 5; a, b, d, f, h, i, and j independently varyfrom 1-10; c, e, g, and k independently vary from 1-100; R_(a), R_(b),R_(c), and R_(d) are defined in the same manner as Y₄; and T is either Hor a negative charge.
 5. The composition as in claims 1, 2, 3, or 4further comprising a contrast agent.
 6. The composition as in claims 1,2, 3, or 4 wherein the compound comprises a radioactive halogen.
 7. Thecomposition as in claims 1, 2, 3, or 4 wherein at least one R group ofthe compound is replaced by a polyamino carboxylic acid or itsderivative.
 8. The composition of claim 7 further comprising aradioactive metal ion or a paramagnetic metal ion.
 9. The composition asin claims 1, 2, 3, 4, 6, or 7 formulated as at least one of a liposome,a micell, a microcapsule, or a microparticle.
 10. The composition as inclaims 1, 2, 3, 4, 6, or 7 formulated as at least one of ultra smalliron oxide particles, silver particles, or gold particles.
 11. A methodfor performing a diagnostic or therapeutic procedure comprisingadministering to a mammal an effective amount of the compound of formula4

wherein at least one of W₄ and X₄ is —CR_(c)R_(d) and the other isselected from the group consisting of —CR_(c)R_(d), —NR_(c), —O—, and—S—; R₂₄, R₂₅, R₂₆, R₂₇, R₂₈, R₂₉, R₃₀, R₃₁, R₃₂, R₃₃, R₃₄, R₃₅ and R₃₆,Y₄, and Z₄ are independently selected from the group consisting ofC1-C10 alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20, polyhydroxyalkyl, C5-C20polyhydroxyaryl, glucose derivatives of R groups, saccharides, amino,C1-C10 aminoalkyl, cyano, nitro, halogen, hydrophilic peptides,arylpolysulfonates, C1-C10 alkyl, C5-C20 aryl, —SO₃T, —CO₂T, —OH,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,—(CH₂)_(a)OCON H(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CON H(CH₂)_(b)PO₃HT,—(CH₂)_(a)CON H(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCON H(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)C—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b))—CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; V₄ is a single bond oris selected from the group consisting of —O—, —S—, —Se—, and —NR_(a); a₄and b₄ vary from 0 to 5; a, b, d, f, h, i, and j independently vary from1-10; c, e, g, and k independently vary from 1-100; R_(a), R_(b), R_(c),and R_(d) are defined in the same manner as Y₄; and T is either H or anegative charge, and thereafter performing the diagnostic or therapeuticprocedure.
 12. A method for performing a diagnostic or therapeuticprocedure comprising administering to a mammal an effective amount ofthe compound of formula 4

wherein at least one of W₄ and X₄ is 0 and the other is selected fromthe group consisting of —CR_(c)R_(d), —NR_(c), —O—, and —S—; R₂₄, R₂₅,R₂₆, R₂₇, R₂₈, R₂₉, R₃₀, R₃₁, R₃₂, R₃₃, R₃₄, R₃₅ and R₃₆, Y₄, and Z₄ areindependently selected from the group consisting of C1-C10 alkoxyl,C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl,glucose derivatives of R groups, saccharides, amino, C1-C10 aminoalkyl,cyano, nitro, halogen, hydrophilic peptides, arylpolysulfonates, C1-C10alkyl, C5-C20 aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —(CH₂)_(a)CONH(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCO(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T, —(CH₂)_(a)OCONH(CH₂)_(b)SO₃T,—(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂, —(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂,—(CH₂)_(a)NHPO₃HT, —(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCON H(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCON H(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b)) —CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; V₄ is a single bondor is selected from the group consisting of —O—, —S—, —Se—, and —NR_(a);a₄ and b₄ vary from 0 to 5; a, b, d, f, h, i, and j independently varyfrom 1-10; c, e, g, and k independently vary from 1-100; R_(a), R_(b),R_(c), and R_(d) are defined in the same manner as Y₄; and T is either Hor a negative charge, and thereafter performing the diagnostic ortherapeutic procedure.
 13. A method for performing a diagnostic ortherapeutic procedure comprising administering to a mammal an effectiveamount of the compound of formula 4

wherein at least one of W₄ and X₄ is NR_(c) and the other is selectedfrom the group consisting of —CR_(c)R_(d), —NR_(c), —O—, and —S—; R₂₄,R₂₅, R₂₆, R₂₇, R₂₈, R₂₉, R₃₀, R₃₁, R₃₂, R₃₃, R₃₄, R₃₅ and R₃₆, Y₄, andZ₄ are independently selected from the group consisting of C1-C10alkoxyl, C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20polyhydroxyaryl, glucose derivatives of R groups, saccharides, amino,C1-C10 aminoalkyl, cyano, nitro, halogen, hydrophilic peptides,arylpolysulfonates, C1-C10 alkyl, C5-C20 aryl, —SO₃T, —CO₂T, —OH,—(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T, —(CH₂)_(a)NHSO₃T,—(CH₂)_(a)CO₂(CH₂)_(b)SO₃T, —(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —(CH₂)_(a)CONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCO(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T,(CH₂)_(a)OCON H(CH₂)_(b)SO₃T, —(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂,—(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂, —(CH₂)_(a)NHPO₃HT,—(CH₂)_(a)NHPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CON H(CH₂)_(b)PO₃HT,—(CH₂)_(a)CON H(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)—NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, (CH₂)_(h)—N(R_(a))—(CH₂)_(i)—CO₂T, and—(CH₂)_(j)—N(R_(b)) —CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; V₄ is a single bondor is selected from the group consisting of —O—, —S—, —Se—, and —NR_(a);a₄ and b₄ vary from 0 to 5; a, b, d, f, h, i, and j independently varyfrom 1-10; c, e, g, and k independently vary from 1-100; R_(a), R_(b),R_(c), and R_(d) are defined in the same manner as Y₄; and T is either Hor a negative charge, and thereafter performing the diagnostic ortherapeutic procedure.
 14. A method for performing a diagnostic ortherapeutic procedure comprising administering to a mammal an effectiveamount of the compound of formula 4

wherein at least one of W₄ and X₄ is S and the other is selected fromthe group consisting of-CR_(c)R_(d), —NR_(c), —O—, and —S—; R₂₄, R₂₅,R₂₆, R₂₇, R₂₈, R₂₉, R₃₀, R₃₁, R₃₂, R₃₃, R₃₄, R₃₅ and R₃₆, Y₄, and Z₄ areindependently selected from the group consisting of C1-C10 alkoxyl,C1-C10 polyalkoxyalkyl, C1-C20 polyhydroxyalkyl, C5-C20 polyhydroxyaryl,glucose derivatives of R groups, saccharides, amino, C1-C10 aminoalkyl,cyano, nitro, halogen, hydrophilic peptides, arylpolysulfonates, C1-C10alkyl, C₅-C₂₀ aryl, —SO₃T, —CO₂T, —OH, —(CH₂)_(a)SO₃T, —(CH₂)_(a)OSO₃T,—(CH₂)_(a)NHSO₃T, —(CH₂)_(a)CO₂(CH₂)_(b)SO₃T,—(CH₂)_(a)OCO(CH₂)_(b)SO₃T, —(CH₂)_(a)CONH(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCO(CH₂)_(b)SO₃T, —(CH₂)_(a)NHCONH(CH₂)_(b)SO₃T,—(CH₂)_(a)NHCSNH(CH₂)_(b)SO₃T, —(CH₂)_(a)OCONH(CH₂)_(b)SO₃T,—(CH₂)_(a)PO₃HT, —(CH₂)_(a)PO₃T₂, —(CH₂)_(a)OPO₃HT, —(CH₂)_(a)OPO₃T₂,—(CH₂)_(a)NHPO₃HT, —(CH₂)_(a)N HPO₃T₂, —(CH₂)_(a)CO₂(CH₂)_(b)PO₃HT,—(CH₂)_(a)CO₂(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)OCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)CONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)CONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCO(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCO(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCONH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCONH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃HT,—(CH₂)_(a)NHCSNH(CH₂)_(b)PO₃T₂, —(CH₂)_(a)OCONH(CH₂)_(b)PO₃HT, and—(CH₂)_(a)OCONH(CH₂)_(b)PO₃T₂, —CH₂(CH₂—O—CH₂)_(c)—CH₂—OH,—(CH₂)_(d)—CO₂T, —CH₂—(CH₂—O—CH₂)_(e)—CH₂—CO₂T, —(CH₂)_(f)—NH₂,—CH₂—(CH₂—O—CH₂)_(g)—CH₂—NH₂, —(CH₂)_(h)—N(R_(a))—(CH₂)₁—CO₂T, and—(CH₂)_(j)—N(R_(b)) —CH₂—(CH₂—O—CH₂)_(k)—CH₂—CO₂T; V₄ is a single bondor is selected from the group consisting of —O—, —S—, —Se—, and —NR_(a),a₄ and b₄ vary from 0 to 5; a, b, d, f, h, i, and j independently varyfrom 1-10; c, e, g, and k independently vary from 1-100; R_(a), R_(b),R_(c), and R_(d) are defined in the same manner as Y₄; and T is either Hor a negative charge, and thereafter performing the diagnostic ortherapeutic procedure.
 15. The method as in claims 11, 12, 13, or 14wherein said procedure utilizes light of wavelength in the region of350-1300 nm.
 16. The method of claim 15 wherein said procedure comprisesmonitoring a blood clearance profile by fluorescence using light ofwavelength in the region of 350 nm to 1300 nm.
 17. The method as inclaims 11, 12, 13, or 14 wherein said procedure comprises monitoring ablood clearance profile by absorption using light of wavelength in theregion of 350 nm to 1300 nm.
 18. The method as in claims 11, 12, 13, or14 wherein the compound contains a radioactive halogen and imaging themammal by at least one of optical imaging and nuclear imaging.
 19. Themethod as in claims 11, 12, 13, or 14 where the compound administeredhas at least one R group replaced by a polyamino carboxylic acid or itsderivative.
 20. The method as in claims 11, 12, 13, or 14 wherein thecompound administered further comprises a radioactive metal ion or aparamagnetic metal ion.
 21. The method as in claims 11, 12, 13, 14, 19,or 20 further comprising imaging by at least one of optical imaging,nuclear imaging, or magnetic resonance imaging.
 22. The method as inclaims 11, 12, 13, 14, or 19 wherein the compound is administered in aformulation selected from at least one of liposomes, micelles,microcapsules, or microparticles.
 23. The method as in claims 11, 12,13, 14, 18, 19, or 20 wherein the compound is administered in aformulation selected from at least one of ultra small iron oxideparticles, silver particles, or gold particles.
 24. The method as inclaims 11, 12, 13, 14, 18, 19, 20, 21, 22, or 23 further comprisingadministering a non-optical contrast agent and imaging by at least oneof magnetic resonance, ultrasound, x-ray, positron emission tomography,computed tomography, optoacoustic imaging, and single photon emissioncomputed tomography.
 25. The method as in claims 11, 12, 13, 14, 18, 19,20, 21, 22, or 23 wherein said procedure is for physiological functionmonitoring.
 26. The method as in claims 11, 12, 13, 14, 18, 19, 20, 21,22, or 23 wherein said procedure is for at least one of renal functionmonitoring, cardiac function monitoring, and kidney function monitoring.27. The method as in claims 11, 12, 13, 14, 18, 19, 20, 21, 22, or 23wherein said procedure is for determining organ perfusion in vivo. 28.The method as in claims 11, 12, 13, 14, 18, 19, 20, 21, 22, or 23further comprising optically imaging the mammal.
 29. The method ofimaging a patient comprising administering a non-optical contrast agentcomposition further comprising the compound as in claims 1, 2, 3, 4, 7,or 8 and performing at least one of an optical imaging procedure or anon-optical imaging procedure.
 30. The method of claim 29 wherein thenon-optical contrast agent composition is chosen from a magneticresonance composition, a computed tomography composition, an x-raycomposition, a nuclear imaging composition, a positron emissiontomography composition, a single photon emission computed tomographycomposition, an optoacoustic imaging composition and an ultrasoundcomposition.
 31. The method of claim 29 wherein the compound stabilizesor buffers the non-optical contrast agent composition.